diethyl 1-(2,4-dichlorophenyl)-5-methyl-4,5-dihydro-1H-pyrazole-3,5-dicarboxylate

Toxicity to birds

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to birds: acute oral toxicity test

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPP 71-1 (Avian Acute Oral Toxicity Test)

GLP compliance:

yes

Dose method:

capsule

Analytical monitoring:

not specified

Test organisms (species):

Coturnix coturnix japonica

Details on test organisms:

TEST ORGANISM
- Common name: Japanese quail (Coturnix coturnix japonica)
- Source: Gudrun Pottiez, Raiffeisenstrasse, D-7519 Eppingen-Adelshofen, Federal Republic of Germany
- Age at test initiation: approx. 6 months
- Weight at test initiation:
male: 164 g (139 g -191 g )
female: 186 g (167 - 210 g)



- Sexes used: 5 male and 5 female
- Cultural background:
- Disease free: yes
- Kept according to standard practices: yes / no
- Breeding population (e.g. reproductive history):

Limit test:

yes

Total exposure duration (if not single dose):

14 d

No. of animals per sex per dose and/or stage:

5 male and 5 female

Control animals:

other: No concurrent control group was used in this study. In the spare group of the same batch of birds as used for the study no mortality or clinical signs or other abnormalities could be observed during the period of this study.

Nominal and measured doses / concentrations:

0 and 2000 mg / kg body weight. Doses were administered in gelatine capsules into the stomach.

Details on test conditions:

ACCLIMATION
- Acclimation period: 14 d
- Health (any disease or mortality observed): No disease or mortality observed

Diet:
During the study the birds were given a special diet (quail grower ration prepared
by the Baierseich Experimental Farm of HOECHST AG. Food and water were available ad libitum.


NO. OF BIRDS PER REPLICATE
- For negative control: 10 (5 male and 5 female)
- For treated: 10 (5 male and 5 female)


NO. OF REPLICATES PER GROUP
- For negative control: 1
- For treated: 1


TEST CONDITIONS (range, mean, SD as applicable)
- Temperature:
- Room temperature: 18 - 21°C
- Relative humidity (%): 40 - 60%.
- Photoperiod: 8 hours light and 16 hours darkness
- Ventilation: yes

Reference substance (positive control):

not specified

Duration (if not single dose):

15 d

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Duration (if not single dose):

15 d

Dose descriptor:

other: LLD (lowest lethal dose)

Effect level:

> 2 000 mg/kg bw

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Findings:

There were no mortalities or clinical signs of intoxication in any testing group. The food consumption body weights of the female birds treated at 2000 mg kg-1 were slightly decreased from days 1-4. consumption and body weights of the other birds appeared not to be affected by the test substance. examination of the birds killed at the end of the observation period revealed no substance-related The LD50 as well as the lowest lethal dose (LLD) was greater than 2000 mg /kg body weight.

Conclusion: The LD50 as well as the lowest lethal dose (LLD) was greater than 2000 mg/kg body weight.

Reference 2

Endpoint:

short-term toxicity to birds: acute oral toxicity test

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPP 71-1 (Avian Acute Oral Toxicity Test)

GLP compliance:

yes

Dose method:

capsule

Analytical monitoring:

not specified

Vehicle:

no

Test organisms (species):

Anas platyrhynchos

Details on test organisms:

TEST ORGANISM
- Common name: Mallard
- Source: Bernhard Franzsander, Heustrasse 15, Delbruck-Hagen, Federal Republic of Germany
- Age at test initiation: approx. 12 months
- Weight at test initiation : male = 1063 g; female = 1065 g
- Sexes used: 5 males and 5 females

Limit test:

yes

Total exposure duration (if not single dose):

15 d

No. of animals per sex per dose and/or stage:

5 males and 5 females

Nominal and measured doses / concentrations:

Nominal: 0 and 2000 mg kg-1

Details on test conditions:

ACCLIMATION
- Acclimation period: 14 d


NO. OF BIRDS PER REPLICATE
- For negative control: 10
- For treated: 10


NO. OF REPLICATES PER GROUP
- For negative control: 1
- For treated: 1


TEST CONDITIONS (range, mean, SD as applicable)
- Temperature:
- Room temperature: 18 - 24°C
- Relative humidity (%): 55 - 80 %
- Photoperiod: lighting cycle of 8 hours light and 16 hours darkness
- Ventilation: yes

Reference substance (positive control):

not specified

Duration (if not single dose):

15 d

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Duration (if not single dose):

15 d

Dose descriptor:

other: LLD ( lowest lethal dose)

Effect level:

> 2 000 mg/kg bw

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Findings:

There were no mortalities or clinical signs of intoxication or impairment of body weight in any testing group. The food consumption of the birds treated at 2000 mg/kg body weight was slightly decreased from days 1-4. The food consumption of the other birds appeared not to be affected. Macroscopic examination of the birds killed at the end of the observation period revealed no substance-related changes. The LD50 as well as the lowest lethal dose (LLD) was greater than 2000 mg/kg body weight.

Conclusion:

The LD50 as well as the lowest lethal dose (LLD) was greater than 2000 mg/kg body weight

Reference 3

Endpoint:

short-term toxicity to birds: dietary toxicity test

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPP 71-2 (Avian Dietary Toxicity Test)

GLP compliance:

yes

Dose method:

feed

Analytical monitoring:

yes

Vehicle:

yes

Details on preparation and analysis of diet:

DIET PREPARATION
- Preparation of doses: A 1 kg finalmixture for each of the 3 dietary concentrations with the test substance and one vehicle control mixture was prepared using sesame oil as mediator according to SOP 049-0/85 and delivered by the Analytical Laboratory of HOECHST AG .
- Type and function of solvent/vehicle: sesame oil was used as the mediator for inclusion of the test substance into the diet

Test organisms (species):

Coturnix coturnix japonica

Details on test organisms:

TEST ORGANISM
- Common name: Japanese quail
- Source:
1) Breeder: HOECHST AG, Pharma Development Toxicology;
2) Source of eggs: Gudrun Pottiez, Raiffeisenstrasse, Eppingen/Adelshofen Federal Republic of Germany
- Age at test initiation: 10 + 1 days at the start of treatment
- Weight at test initiation: 28.2 g
- Sexes used: males and females (sexing phenotypically impossible in birds of this age)

Limit test:

no

Total exposure duration (if not single dose):

8 d

Remarks:

last 3 days untreated diet

No. of animals per sex per dose and/or stage:

males and females (sexing phenotypically impossible in birds of this age)

Control animals:

yes

Nominal and measured doses / concentrations:

Nominal concentrations of 625, 1750 and 5000 ppm test item

Details on test conditions:

ACCLIMATION
- Acclimation period: 3 days
- Feeding: Food and water were available ad libitum.
- Health (any disease or mortality observed):


NO. OF BIRDS PER REPLICATE
- For negative control: 10
- For treated: 10
- For vehicle control: 10

NO. OF REPLICATES PER GROUP
- For negative control: 1
- For treated: 1
- For vehicle control: 1


TEST CONDITIONS (range, mean, SD as applicable)
- Temperature: The temperature in the rearing cages was about 36 °C in the 1st week after hatching, 32 °C in the 2nd week, and 28 °C in the 3rd week
- Relative humidity (%): 40 %
- Photoperiod: lighting cycle of 16 hours light and 8 hours darkness

Details on examinations and observations:

During the treatment and observation periods, the signs of intoxication and the mortality rate were recorded twice daily. The birds were weighed at the start of the study (day -3), at the start of treatment (day 1 = day of dosing) and during the observation period on days 6 and 9. The food consumption of these groups was also recorded for days 1-6 (5-day treatment period) and 6-9 (3- day treatment-free period). At the end of the observation period the surviving animals were killed by carbon dioxide asphyxiation, dissected and macroscopically examined.

Reference substance (positive control):

not specified

Duration (if not single dose):

5 d

Dose descriptor:

NOEL

Effect level:

> 5 000 mg/kg diet

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Duration (if not single dose):

5 d

Dose descriptor:

NOEL

Effect level:

ca. 1 462 mg/kg bw

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Findings:

Analysis of diets for AE F107892 (content, homogeneity, stability) was carried out and revealed actual diet concentrations (mean values) of 98 - 108 % of the nominal diet concentrations. No mortality, clinical signs of toxicity or effects on either food consumption or body weight were observed in any treated group up to and including 5000 ppm. Macroscopic examination of the birds killed at the end of the observation period revealed no substance-related changes. The LC50 , the LLC (lowest lethal concentration), and the NOAEL (No Observed Adverse Effect level) were thus greater than 5000 mg/kg diet, equivalent to a mean daily substance intake of ca. 1462 mg/kg body weight 1.

Conclusion:

The LC50 , the LLC (lowest lethal concentration), and the NOAEL (No Observed Adverse Effect level) were thus greater than 5000 mg/kg diet, equivalent to a mean daily substance intake of ca. 1462 mg/kg body weight.

Reference 4

Endpoint:

short-term toxicity to birds: dietary toxicity test

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPP 71-2 (Avian Dietary Toxicity Test)

GLP compliance:

yes

Dose method:

feed

Analytical monitoring:

yes

Vehicle:

yes

Test organisms (species):

Anas platyrhynchos

Details on test organisms:

TEST ORGANISM
- Common name: Mallard ducks
- Source: B. Franzsander, D-6369 Schöneck OT Oberdorfelden, FalltorstraBe 21, Federal Republic of Germany
- Age at test initiation:5 days
- Weight at test initiation: 81.1 g
- Sexes used: males and females (sexing phenotypically impossible in birds of this age)

Limit test:

no

Total exposure duration (if not single dose):

8 d

No. of animals per sex per dose and/or stage:

Each group contained 10 birds, randomly selected, without regard to sex.

Control animals:

yes

Nominal and measured doses / concentrations:

Nominal concentrations of 312.5, 625, 1250, 2500 and 5000 ppm AE F107892

Details on test conditions:

ACCLIMATION
- Acclimation period: 3 days
- Feeding: Food and water were available ad libitum (special chick starter ration manufactured by Altromin GmbH).
- Health: yes

NO. OF BIRDS PER REPLICATE
- For negative control: 10
- For vehicle control: 10
- For treated: 10

NO. OF REPLICATES PER GROUP
- For negative control: 1
- For vehicle control: 1
- For treated: 1

TEST CONDITIONS (range, mean, SD as applicable)
- Temperature: 21 - 23 °C
- Relative humidity (%): 50 - 60 %
- Photoperiod: 17 hours light / 7 hours darkness (17L:7D) light intensity in cages 100 - 300 lux)

Reference substance (positive control):

not specified

Duration (if not single dose):

5 d

Dose descriptor:

NOEL

Effect level:

> 5 000 mg/kg diet

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Duration (if not single dose):

5 d

Dose descriptor:

NOEL

Effect level:

> 1 590 mg/kg bw

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Findings:

Analysis of diets for AE F107892 (content, homogeneity, stability) was carried out and revealed actual diet

concentrations (mean values) of 93 - 106 % of the nominal diet concentrations.

No mortality*, clinical signs of toxicity or effects on either food consumption or body weight were observed in

any treated group up to and including 5000 ppm. Macroscopic examination of the birds killed at the end of the

observation period revealed no substance-related changes. The LC50 , the LLC (lowest lethal concentration), and

the NOAEL (No Observed Adverse Effect level) were thus greater than 5000 mg/kg diet, equivalent to a mean

daily substance intake of ca. 1590 mg/kg body weight.

*During the study period one duckling in the 1250 ppm group and one in the 2500 ppm group died on days 4 and

7, respectively. Both deaths were considered to be incidental, since no mortality occurred in the highest dose

group (5000 ppm).

Conclusion:

The LC50, the LLC (lowest lethal concentration), and the NOAEL (No Observed Adverse Effect level) were

greater than 5000 mg/kg diet, equivalent to a mean daily substance intake of ca. 1590 mg/kg body weight.

Reference 5

Endpoint:

long-term toxicity to birds: reproduction test

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 206 (Avian Reproduction Test)

Deviations:

yes

Remarks:

Appropriate for non-bioaccumulating substances. As compared with TG 206 prelaying phase was omitted and 8-wk laying period reduced to 6-wk due to higher sensitivity of new study design including a 2-wk pre-study phase for taking out subfertile replicates.

GLP compliance:

yes

Dose method:

feed

Analytical monitoring:

yes

Vehicle:

no

Details on preparation and analysis of diet:

DIET PREPARATION
- Preparation of doses: For each test concentration excluding the control, 1 kg premix containing a concentration 25 times (1st batch) or 15 times (2nd batch) higher than the final dietary concentration was delivered by the ANALYTICAL LABORATORY. The premixes were produced without a mediator substance. Each premix (1 kg) was mixed with 14 or 24 kg diet (total 15 or 25 kg) in two stages for approx. 30 minutes, and stored in closed plastic containers in the testing room at room temperature.

Test organisms (species):

Coturnix coturnix japonica

Details on test organisms:

TEST ORGANISM
- Common name: Japanese quails
- Source: Hoechst Aktiengesellschaft Corporate Toxicology
- Age at test initiation: approx. 12 weeks
- Weight at test initiation:
cocks= 190 g
hens= 221 g
- Sexes used: 64 cocks / 64 hens

Limit test:

no

Total exposure duration (if not single dose):

6 wk

No. of animals per sex per dose and/or stage:

parent animals: 16 per sex per dose

Control animals:

yes, concurrent no treatment

Nominal and measured doses / concentrations:

Nominal concentrations: 0, 40, 200 or 1000 ppm diet, which were equivalent to a mean daily substance intake of 0, 4.2, 21.6 and 106 mg kg-1, respectively.

Details on test conditions:

ACCLIMATION
- Acclimation period: 1 week
- Acclimation conditions (same as test or not): same
- Feeding: ad libitum
- Health: yes


NO. OF BIRDS PER REPLICATE
- 64 quail pairs (16 pairs)


NO. OF REPLICATES PER GROUP
- For negative control: 1
- For treated: 1


TEST CONDITIONS (range, mean, SD as applicable)
- Temperature: 21 - 23 °C
- Relative humidity (%): 50 - 60 %
- Photoperiod: 17 hours light / 7 hours darkness (17L:7D)
light intensity in cages 100 - 300 lux)

Details on reproductive parameters:

The following parameters were examined:
- Eggs laid
- Eggs cracked; eggs broken
- Egg abnormalities
- Eggs set
- Eggshell thickness or eggshell strength
- Eggs fertile
- Embryos viable
- Normal hatchlings
- Abnormal hatchlings
- Clinical signs of toxicity, abnormalities and mortality
- 14-day old surviving chicks
- Chick body weight at hatching and 14 days after hatching

Reference substance (positive control):

not specified

Duration (if not single dose):

6 wk

Dose descriptor:

NOEC

Effect level:

>= 1 000 mg/kg diet

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Remarks:

and reproduction

Duration (if not single dose):

6 wk

Dose descriptor:

NOEC

Effect level:

>= 106 mg/kg bw

Conc. / dose based on:

test mat.

Basis for effect:

mortality

Remarks:

and reproduction

Findings:

Analysis of diets for AE F107892 (content, homogeneity, stability) were conducted and revealed actual diet

concentrations of 100 - 107 % of the nominal diet concentrations.

The behaviour, general health condition and food consumption of the pairs in all groups treated up to and

including 1000 ppm proved to be unaffected. Necropsy indicated no changes either in the organs or in the organ

weight analysis of heart, liver, spleen, testes and oviduct. No impairment of egg production was detected in any

group. The mean egg weight, the numbers of broken/cracked eggs and abnormal eggs as well as the shell

thickness indicated no substance-related changes. The fertility and hatching rate of the groups treated with the

test substance indicated no substance-related changes. The hatching success for the incubated control eggs was

67 %, thus fulfilling the quality criteria of at least 50 %. The behaviour and general health condition of the

chicks from study weeks 4 to 6 appeared to be normal in all groups. The 14-day survival in all groups remained

unaffected by AE F107892. The survival rate in the control group was 93 % of the hatchlings, thus fulfilling the

quality criterion of > 80 %. The chick weights from all treated groups showed no statistically significant changes

as compared with those of the control at hatching and after completion of the 14-day rearing period.

Mortality, body weight, food consumption and reproduction of Japanese quails after

6-week dietary exposure to AE F107892

Examination	Nominal diet concentration (ppm)
	0	40	200	1000
No. of replicates at study end	16	16	16	15
Replicate body weight mean WK 1 - 6 [g]	415.7	408.0	409.4	408.3
Food consumption WK 1 - 6 [g/day]	42.9	43.3	44.3	43.2
Food consumption [g/100g bw/day]	10.3	10.6	10.8	10.6
Substance intake [mg/kg bw/day]	-	4.2	21.6	106
Eggs laid per hen	32.7	34.1	35.5	35.3
Eggs intact / eggs laid [%]	84.6	93.3	90.7	87.9
Mean egg shell thickness [mm]	0.22	0.22	0.23	0.22
Fertile eggs / eggs set [%]	92.3	94.3	92.4	93.8
Chicks hatched / eggs fertile [%]	73.6	80.6	80.3	66.7
Viable chicks per hen	18.8	24.6	23.6	20.7
14-days survivors / hatched [%]	93.4	92.8	91.4	93.3
Chick weight day 1 [g]	8.9	8.7	8.9	8.6
Chick weight day 14 [g]	66.8	65.4	62.5	65.3

Conclusion:

Based on these results, the "No Observed Effect Concentration" (NOEC) was considered to be 1000 ppm,

equivalent to a mean daily substance intake of 106 mg kg-1. The study provided reliable endpoint data for

assessment of subchronic / reproductive risk to birds.

Comment:

This study provides reliable endpoint data for risk assessment as all validity criteria as set in OECD

guideline 206 have been fulfilled. The scientific approach of this study design can be found in the publication: B.

Schlatterer (1993). Effects of bis (tri-n-butyltin) oxide in the Japanese quail exposed during egg laying period:

An interlaboratory comparison study. Arch. Environ. Contam. Toxicol. 24, 440-448.

Additional information

Five studies investigating the toxicity of the substance to birds are available. The key study was conducted according to OECD guideline 206 (1994). Japanese quails were fed with three different concentrations of the test substance. The mean daily substance intake was 4.2, 21.6 and 106 mg /kg bw, respectively. During the test period of six weeks the reproduction and mortality of the test animals was recorded. Behaviour, health and reproduction were not impaired by the test substance. The determined NOEC (42 d) was ≥ 106 mg/kg bw. The study is supported by two acute oral tests following EPA OPP 71-1 (1991a and 1991 b). The test substance was fed to Japanese quails and Mallard ducks respectively by means of gelatine capsules (dose: 2000 mg/kg bw). During the test period of 14 days no disease or mortality was observed. The LD50 (14 d) was >2000 mg /kg body weight. Two further acute tests, conducted according to EPA OPP 71-2, investigated the oral toxicity of the substance to Japanese quails and the Mallard ducks (1991c and 1992). The test substance was fed to the test animals over a period of 5 days, followed by a period of 3 days with untreated food. Neither mortality nor clinical signs of intoxication attributable to the test substance were observed in any study. The determined NOEL (5 d) in both studies was > 5000 mg/kg diet based on mortality.  

The substance has a low potential for short- and long-term toxicity to birds.